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Abstracts: CryoLetters 29 (1), 2008

CryoLetters 29(1), 1-6 (2008)
© CryoLetters, c/o Royal Veterinary College, London NW1 0TU, UK

 CHRIS POLGE, CBE FRS - (16 AUGUST 1926 – 17 AUGUST 2006)
A PERSONAL APPRECIATION

Paul Watson

The Royal Veterinary College, London Royal College Street, NW1 0TU, UK (pwatson@rvc.ac.uk)

 

 

CryoLetters 29(1), 7-14 (2008)
© CryoLetters, c/o Royal Veterinary College, London NW1 0TU, UK

DETERMINATION OF GENETIC STABILITY IN SURVIVING APPLE SHOOTS FOLLOWING CRYOPRESERVATION BY VITRIFICATION

Yun-Guo Liu 1, 2*, Ling-Xiao Liu 3, Lei Wang1, Ai-Ying Gao4

1 Division of Life Science and Technology, Ocean University of China, Qingdao, 266003, China
2 Agricultural Product Inspection Center, Shandong Entry-Exit Inspection & Quarantine Bureau, Qingdao 266002, China
3 Linyi Academy of Agricultural Sciences, Linyi 276012, China
4Taian Institute of Supervision & Inspection on Product Quality, Taian 271000, China
*Corresponding author: Yun-Guo Liu,
Tel: 86-532-86770612 E-mail:
yguoliu@163.com

Abstract

In vitro-grown apple shoot tips were successfully cryopreserved by vitrification, with an average survival and shoot formation of 80% and 76% respectively. Surviving shoots showed the same rate and  regrowth patterns as those of non-treated controls. No significant differences (P<0.05) were observed in morphological characteristics, including shoot length, leaf shape, leaf width/length ratio and root length, between the control and cryopreserved shoots. No different microsatellite alleles and ISSR fragments were detected between control and cryopreserved shoots using twelve pairs of microsatellites and eleven ISSR primers. These results show that cryopreservation using vitrification is a practical method for the long-term storage of apple germplasm.

Keywords: genetic stability, apple, vitrification, cryopreservation, microsatellite, ISSR

 

 

CryoLetters 29(1), 15-20 (2008)
© CryoLetters, c/o Royal Veterinary College, London NW1 0TU, UK

DEPLOYMENT OF THE ENCAPSULATION/DEHYDRATION PROTOCOL TO CRYOPRESERVE MICROALGAE HELD AT THE SAMMLUNG VON ALGENKULTUREN, UNIVERSITÄT GÖTTINGEN, GERMANY

Keith Harding1,2*, Julia Müller3, Maike Lorenz3, Hella Timmerman3,
Thomas Friedl3, John G. Day4 & Erica E. Benson1,2

1Plant Conservation Group, University of Abertay Dundee, Bell Street, Dundee, DD1 1HG, UK
2*Current address for correspondence: Damar Research Scientists, Conservation, Environmental Science and Biotechnology, Drum Road, Cuparmuir, Cupar, Fife, KY15 5RJ, Scotland UK. E-mail: 
k.harding-damar@tiscali.co.uk
3Albrecht-von-Haller-Institut für Pflanzenwissenschaften, Abteilung Experimentelle Phykologie und Sammlung von Algenkulturen, Universität Göttingen, Untere Karspüle 2, 37073 Göttingen, Germany
4Culture Collection of Algae and Protozoa, Scottish Association for Marine Science, Dunstaffnage Marine Laboratory, Dunbeg, Argyll, PA37 1QA, Scotland UK.

Abstract

Encapsulation/dehydration was applied to cryopreserve 14 diverse algal strains, representing eukaryotic terrestrial microalgae; of these 12 survived to form cell colonies after recovery from cryostorage. Surviving algae had varying degrees of tolerance to osmotic dehydration and desiccation in this vitrification-based cryoprotective strategy. The extent of algal regrowth was affected by the mode of desiccation (silica gel or air-flow), the duration of evaporative desiccation and exposure to light during early recovery phase. This paper: (i) demonstrates the versatility of the encapsulation/dehydration method to cryopreserve diverse microalgae; (ii) confirms the successful transfer of this cryostorage technology to the Culture Collection of Algae at Göttingen University (SAG); and (iii) recommends encapsulation/dehydration as a feasible alternative to controlled rate cooling for preserving algae held in international culture collections.

Keywords: culture collections, cryopreservation, encapsulation, microalgae, vitrification

 

 

CryoLetters 29(1), 21-26 (2008)
© CryoLetters, c/o Royal Veterinary College, London NW1 0TU, UK

DEPLOYMENT OF THE ENCAPSULATION/DEHYDRATION PROTOCOL TO CRYOPRESERVE DIVERSE MICROALGAE HELD AT THE INSTITUTE OF SOIL BIOLOGY, ACADEMY OF SCIENCES OF THE CZECH  REPUBLIC

Alena Lukešová1*, Pavel Hrouzek1, Keith Harding2,3, Erica E Benson2,3 & John G Day4

1Institute of Soil Biology, Biology Centre AS CR, v.v.i., Na Sádkach 7, 370 05, Čské Buděovice, Czech Republic
*Corresponding author - Email:
luksa@upb.cas.cz
2University of Abertay Dundee, Bell Street, Dundee, DD1 1HG, UK
3Current address: Damar, Research Scientists, Drum Road, Cupar Muir, Fife, KY15 5RJ, UK
4Culture Collection of Algae and Protozoa, Scottish Association for Marine Science, Dunstaffnage Marine Laboratory, Dunbeg, Argyll, PA37 1QA, UK

Abstract

Twenty-seven strains of soil algae isolated from highly diverse provenances and habitats were assessed for their capacity to withstand cryopreservation using encapsulation/dehydration. Survival was assessed following the release of algae from alginate beads treated with sodium hexametaphosphate and regrowth was assessed using NAJA® Image Analysis. Regrowth occurred in 19 strains, with >50% survival being observed in 15. Algal tolerance to osmotic dehydration and evaporative desiccation was critical to the success of the method. Recovery in five out of the remaining eight recalcitrant strains was enhanced by substituting sorbitol for the osmotic pretreatment or by combining encapsulation with two-step controlled rate cooling.

Keywords: cryopreservation, microalgae, cyanobacteria, culture collections, vitrification

 

 

CryoLetters 29(1), 27-28 (2008)
© CryoLetters, c/o Royal Veterinary College, London NW1 0TU, UK

DEPLOYMENT OF THE ENCAPSULATION/DEHYDRATION PROTOCOL TO CRYOPRESERVE POLAR MICROALGAE HELD AT THE CZECH REPUBLIC ACADEMY OF SCIENCES INSTITUTE OF BOTANY

Josef Elster1*, Jaromír Lukavský1, Keith Harding2,3, Erica E. Benson2,3 and John G. Day4

1*Inst. Botany AS CR, Dukelska 135, CZ 379 83 Třboň Czech Republic.
Email:
jelster@butbn.cas.cz
2Plant Conservation Group, University of Abertay Dundee, Dundee, DD1 1HG, UK.
3Current address: Research Scientists, Damar, Drum Road, Cuparmuir, Fife, KY15 5RJ, UK.
4Culture Collection of Algae and Protozoa, SAMS, Dunbeg, Argyll, PA37 1QA, UK.

Abstract

Polar isolates of four chlorococcal microalgae originating from the Arctic and Antarctica withstand cryopreservation using encapsulation/dehydration. Viability assessments, which initially used chloroplhyll fluorescence (Kautsky) induction kinetics, revealed that all strains suffered photosynthetic impairment during early post-cryopreservation recovery. This cryoinjury was reversible, as indicated by cell regrowth in three of the four strains. Lack of growth in the fourth isolate was due to contaminating bacteria rather than cryogenic factors.

Keywords: polar biology, cryopreservation, encapsulation, vitrification

 

 

CryoLetters 29(1), 29-34 (2008)
© CryoLetters, c/o Royal Veterinary College, London NW1 0TU, UK

CUBIC ICE CAN BE FORMED DIRECTLY IN THE WATER PHASE OF VITRIFIED AQUEOUS SOLUTIONS

Kazuhito Kajiwara1, Pariya Thanatuksom1, Norio Murase2 and Felix Franks3*

1School of Bionics, Tokyo University of Technology, 1404-1 Katakura, Hachioji, Tokyo, 192-0892 Japan
2Department of Biotechnology, Tokyo Denki University, Hatoyama-cho, Hiki-gun, Saitama, 350-0394, Japan
3BioUpdate Foundation, 25 The Fountains, 229 Ballards Lane, London N3 1NL, United Kingdom
*Corresponding author: *E-mail:
bioup@dial.pipex.com

Abstract

This study addresses the possibility that ice-Ic may form spontaneously and directly during the cooling of previously freeze-concentrated aqueous solutions, at temperatures where the residual unfrozen water is likely to exist in small, isolated domains. It is shown that on cooling a dilute aqueous glucose solution, ice-Ih crystallises initially, but when the solution has been freeze-concentrated to ca.10 mol%, further cooling results in the slow nucleation and growth of ice-Ic in the residual water domains, rather than on the pre-existing ice-Ih crystal surfaces.

Keywords: Cubic ice, freeze concentration of glucose, glass transition, thermal analysis, X-ray diffraction, ice polymorphism.

 

 

CryoLetters 29(1), 35-42 (2008)
© CryoLetters, c/o Royal Veterinary College, London NW1 0TU, UK

INC REASE IN THE CONTRIBUTION OF TRANSAMINATION TO THE RESPIRATION OF MITOCHONDRIA DURING AROUSAL

N.I.Fedotcheva1*, E.G.Litvinova1,  Z.G.Amerkhanov2, S.V.Kamzolova3, I.G.Morgunov3 and  M.N.Kondrashova 1

1Institute of Theoretical and Experimental Biophysics, Russian Academy of Sciences, Pushchino, Moscow region, 142290 Russia
2Institute of Cell Biophysics, Russian Academy of Sciences, Pushchino, Moscow region, 142290 Russia
3Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Pushchino, Moscow region, 142290 Russia
*Corresponding author: NI Fedocheva  E-mail :
nfedotcheva@mail.ru

Abstract

The involvement of transamination in the respiration of liver mitochondria in ground squirrels during hibernation and arousal has been studied. It was shown by HPLC that, in the presence of glutamate and malate, the formation of alpha-ketoglutarate (KGL), a transamination marker, and fumarate, a product of succinate oxidation, takes place. During arousal, the formation of KGL increased fourfold, and the respiration sensitive to the inhibitor of aspartate transaminase aminooxyacetate (AOA) increased threefold. The function of transamination upon arousal is related to the elimination of oxaloacetate, an endogenous inhibitor of succinate dehydrogenase. In addition, being more resistant to oxidative stress than oxidation, transamination is probably involved in the antioxidant defense required during the rapid rise of body temperature upon arousal. Our experiments showed an increase in the concentration of malonic dialdehyde (MDA), an end product of lipid peroxidation, in liver mitochondria in this state, which can reduce the activity of the enzymes of the tricarboxylic acids cycle. Under these conditions, the transamination contributes to the maintenance of a high respiration rate necessary for arousal.

Keywords: arousal, hibernation, HPLC, αketoglutarate, liver, MDA, oxidative stress, transamination

 

 

CryoLetters 29(1), 43-52 (2008)
© CryoLetters, c/o Royal Veterinary College, London NW1 0TU, UK

CRYOBIOLOGY, WILDLIFE CONSERVATION & REALITY

William V. Holt

Institute of Zoology, Zoological Society of London, Regent's Park, London NW1 4RY
Email
Bill.holt@ioz.ac.uk

Abstract

Conservation is about protecting and nurturing species so that they can survive, not only now but also into the future. Ideally this means protecting genetically diverse populations and not simply breeding a few individuals. In principle, cryobiology offers the means to help maintain genetic diversity by storing genetically important germplasm that could reinvigorate populations in the future. Unfortunately the technical problems associated with this ideal goal still provide a major barrier to the practical use of cryopreservation technology. Sometimes these are technical problems with the cryobiology, but lack of basic biological information about unusual species, coupled with difficulties in obtaining such information, means that progress will be possible with only a few species that are subject to intensive scrutiny. The opportunities nevertheless exist for cryobiologists and reproductive biologists to make useful and global contributions to species conservation. I argue here that there are often two mutually suspicious groups of biologists, who do not interact or even understand each others goals. If conservation biologists and biotechnologists were more prepared to join forces and share their expertise, there would be much improved prospects for achieving lasting success in the conservation of a small, but well targeted, number of threatened species.

Keywords: artificial insemination, genetic resource banks, biobanks, cloning.

 

 

CryoLetters 29(1), 53-62 (2008)
© CryoLetters, c/o Royal Veterinary College, London NW1 0TU, UK

CRYOPRESERVATION FOR PLANT GENEBANKS - A MATTER BETWEEN HIGH EXPECTATIONS AND CAUTIOUS RESERVATION

E. R. Joachim Keller*, Anja Kaczmarczyk and Angelika Senula

Genebank Department, Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstrasse 3, D-06466 Gatersleben, Germany.
*Corresponding author: E. R. Joachim Keller.
E-mail:
keller@ipk-gatersleben.de

Abstract

Cryopreservation is the best method of storing germplasm efficiently and safely, particularly for the maintenance of vegetatively propagated material. In IPK cryopreservation is used for potato, garlic, mint and yam. IPK collaborates with other cryobanks and research groups (ECPGR, COST, EURALLIVEG) and finds considerable differences in the adoption of cryopreservation between crops and their host institutes, depending on crop, local and historical circumstances. A better understanding of the long-term benefits of cryopreservation and its further integration into general genebank management is therefore needed. Recommended approaches include: comparative validation of methods between different laboratories, detailed comparisons of crop-based methods, economical analyses, efficient integration strategies of cryobanks by genebanks; including safe duplication of cryopreserved resources for the limitation of risk of loss  Importantly, there has been recent progress in the development of quality management systems. Cryopreservation is, however, characterized by high expectations. Therefore, to ensure its sustainable and practicable use, basic knowledge of storage protocols must be combined with increased awareness of the rationales required to validate, implement and apply cryobanking technologies in working genebanks.

Keywords: genebank management, validation, methodical comparison, cost efficiency, quality management system

 

 

CryoLetters 29(1), 63-72 (2008)
© CryoLetters, c/o Royal Veterinary College, London NW1 0TU, UK

THE IMPACT OF OECD BEST PRACTICE ON THE VALIDATION OF CRYOPRESERVATION TECHNIQUES FOR MICROORGANISMS

David Smith* and Matthew J. Ryan

CABI Europe-UK, Bakeham Lane, Egham, Surrey TW20 9TY UK
*Corresponding author: David Smith  Email:
d.smith@cabi.org

Abstract

The Organisation for Economic Co-operation and Development (OECD) Biological Resource Centre Initiative (BRC) was established after the 1998 Working Party on Biotechnology endorsed a proposal by Japan to examine support for Biological Resource Centres (BRCs) as a key element of the scientific and technological infrastructure for the life sciences and biotechnology. As part of this Best Practice Guidelines for the operation of Biological Resource Centres (BRCs) were published. Cryopreservation is widely used in BRC's and is seen as the method of choice for the preservation of most organism groups. This paper reviews the developments of BRC standards, how they are applied, current practices in cryopreservation and methods for validating the success of cryopreservation methodology.

Keywords: OECD, BRC, Fungi, Bacteria, Yeast, Cryopreservation, Best practice

 

 

CryoLetters 29(1), 73-88 (2008)
© CryoLetters, c/o Royal Veterinary College, London NW1 0TU, UK

ABSTRACTS FROM THE 2007 ANNUAL MEETING OF THE SOCIETY FOR LOW TEMPERATURE BIOLOGY,
UNIVERSITY OF DERBY, UK, 12-14 SEPTEMBER 2007
SYMPOSIUM

"Validation, Safety and Ethical Issues Impacting the Low Temperature Storage of Biological Resources"

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